嗜热病毒的功能生物学和生物技术。
Functional biology and biotechnology of thermophilic viruses.
机构信息
School of Life Sciences, University of Nevada, Las Vegas, Las Vegas, Nevada, U.S.A.
Varizymes, Middleton, Wisconsin, U.S.A.
出版信息
Essays Biochem. 2023 Aug 11;67(4):671-684. doi: 10.1042/EBC20220209.
Viruses have developed sophisticated biochemical and genetic mechanisms to manipulate and exploit their hosts. Enzymes derived from viruses have been essential research tools since the first days of molecular biology. However, most viral enzymes that have been commercialized are derived from a small number of cultivated viruses, which is remarkable considering the extraordinary diversity and abundance of viruses revealed by metagenomic analysis. Given the explosion of new enzymatic reagents derived from thermophilic prokaryotes over the past 40 years, those obtained from thermophilic viruses should be equally potent tools. This review discusses the still-limited state of the art regarding the functional biology and biotechnology of thermophilic viruses with a focus on DNA polymerases, ligases, endolysins, and coat proteins. Functional analysis of DNA polymerases and primase-polymerases from phages infecting Thermus, Aquificaceae, and Nitratiruptor has revealed new clades of enzymes with strong proofreading and reverse transcriptase capabilities. Thermophilic RNA ligase 1 homologs have been characterized from Rhodothermus and Thermus phages, with both commercialized for circularization of single-stranded templates. Endolysins from phages infecting Thermus, Meiothermus, and Geobacillus have shown high stability and unusually broad lytic activity against Gram-negative and Gram-positive bacteria, making them targets for commercialization as antimicrobials. Coat proteins from thermophilic viruses infecting Sulfolobales and Thermus strains have been characterized, with diverse potential applications as molecular shuttles. To gauge the scale of untapped resources for these proteins, we also document over 20,000 genes encoded by uncultivated viral genomes from high-temperature environments that encode DNA polymerase, ligase, endolysin, or coat protein domains.
病毒已经开发出复杂的生化和遗传机制来操纵和利用它们的宿主。自分子生物学的最初阶段以来,来自病毒的酶一直是必不可少的研究工具。然而,已经商业化的大多数病毒酶都来自少数几种培养的病毒,考虑到宏基因组分析揭示的病毒的非凡多样性和丰富性,这是值得注意的。鉴于过去 40 年来源自嗜热原核生物的新型酶试剂的爆炸式增长,源自嗜热病毒的那些试剂也应该是同样有效的工具。
本文综述了嗜热病毒的功能生物学和生物技术的现状,重点讨论了 DNA 聚合酶、连接酶、内切溶素和衣壳蛋白。对感染 Thermus、Aquificaceae 和 Nitratiruptor 的噬菌体的 DNA 聚合酶和引物聚合酶的功能分析揭示了具有强大校对和逆转录酶能力的新酶类。已经从 Rhodothermus 和 Thermus 噬菌体中鉴定出嗜热 RNA 连接酶 1 同源物,其中两种已商业化用于单链模板的环化。来自感染 Thermus、Meiothermus 和 Geobacillus 的噬菌体的内切溶素表现出对革兰氏阴性和革兰氏阳性细菌的高稳定性和异常广泛的裂解活性,使其成为商业化作为抗菌剂的目标。感染 Sulfolobales 和 Thermus 菌株的嗜热病毒的衣壳蛋白已被表征,具有作为分子穿梭的多种潜在应用。为了评估这些蛋白质未开发资源的规模,我们还记录了来自高温环境的未培养病毒基因组编码的超过 20000 个基因,这些基因编码 DNA 聚合酶、连接酶、内切溶素或衣壳蛋白结构域。